SiRAV1蛋白的Ser31位点磷酸化可以通过调节<em>SiCAT</em>表达来提高谷子耐盐性

Journal of Integrative Agriculture ›› 2023, Vol. 22 ›› Issue (12): 3638-3651.DOI: 10.1016/j.jia.2023.04.034

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SiRAV1蛋白的Ser31位点磷酸化可以通过调节SiCAT表达来提高谷子耐盐性

收稿日期:2022-12-22 接受日期:2023-04-04 出版日期:2023-12-20 发布日期:2023-12-06

Phosphorylation of SiRAV1 at Ser31 regulates the SiCAT expression to enhance salt tolerance in Setaria italica

LI Qiao-lu¹, LI Zhi-yong², WANG Meng-meng^3#, YAN Jing-wei^4#, FANG Lin^1#

¹Guangdong Provincial Key Laboratory of Applied Botany, South China Botanical Garden, Chinese Academy of Sciences, Guangzhou 510650, P.R.China

²Institute of Millet Crops, Hebei Academy of Agriculture and Forestry Sciences, Shijiazhuang 050000, P.R.China
³Nanjing Institute of Environmental Science, Ministry of Ecology and Environment, Nanjing 210042, P.R.China
⁴State Key Laboratory of Subtropical Silviculture, Zhejiang A&F University, Hangzhou 311300, P.R.China

Received:2022-12-22 Accepted:2023-04-04 Online:2023-12-20 Published:2023-12-06
About author:LI Qiao-lu, E-mail: l2295397739@163.com; #Correspondence WANG Meng-meng, E-mail: wangmeng2336@163.com; YAN Jing-wei, E-mail: jingweiyan@zafu.edu.cn; FANG Lin, E-mail: linfang@scbg.ac.cn
Supported by:
This work was funded by the National Natural Science Foundation of China (31902062) and the South China Botanical Garden, Chinese Academy of Sciences (QNXM-02).

摘要/Abstract

摘要：

盐害严重威胁着植物的生长和发育。因此，识别和筛选盐胁迫响应的关键基因并解析其作用机制对提高植物的耐盐性至关重要。本研究运用磷酸化蛋白质组技术对盐胁迫处理前后的谷子样品进行检测分析，共鉴定出4000个磷酸化多肽，其中123个显著差异。非冗余蛋白质数据库（NR）功能注释显示，有23个转录因子表达差异显著，如SiRAV1转录因子。亚细胞定位分析显示SiRAV1位于细胞核中。表型和生理分析证实，SiRAV1的过表达能够提高谷子对盐胁迫的耐受性，且这一作用是通过抑制盐诱导的H₂O₂积累、丙二醛含量和电解质渗漏率的增加等方式实现的。进一步的研究表明，SiRAV1能够直接结合SiCAT启动子以激活SiCAT的表达，从而提高CAT酶的活性。本研究还发现，SiRAV1蛋白的Ser31位点磷酸化可以通过增强其与SiCAT启动子的结合能力来正向调节谷子的耐盐性。综上所述，本研究首次解析了SiRAV1在谷子盐响应中的作用机制：盐胁迫通过诱导SiRAV1蛋白的Ser31位点磷酸化，进而增强了其与SiCAT启动子的结合能力，激活了SiCAT的表达，从而提高谷子对盐胁迫的耐受性。

Abstract: Salinity severely affects plant growth and development. Thus, it is crucial to identify the genes functioning in salt stress response and unravel the mechanism by which plants against salt stress. This study used the phosphoproteomic assay and found that 123 of the 4 000 quantitative analyzed phosphopeptides were induced by salt stress. The functional annotation of the non-redundant protein database (NR) showed 23 differentially expressed transcription factors, including a phosphopeptide covering the Serine 31 in the RAV (related to ABI3/VP1) transcription factor (named SiRAV1). SiRAV1 was located in the nucleus. Phenotypic and physiological analysis showed that overexpressing SiRAV1 in foxtail millet enhanced salt tolerance and alleviated the salt-induced increases of H₂O₂ accumulation, malondialdehyde (MDA) content, and percent of electrolyte leakage. Further analysis showed that SiRAV1 positively regulated SiCAT expression to modulate the catalase (CAT) activity by directly binding to the SiCAT promoter in vivo and in vitro. Moreover, we found that phosphorylation of SiRAV1 at the Ser31 site positively regulated salt tolerance in foxtail millet via enhancing its binding ability to SiCAT promoter but did not affect its subcellular localization. Overall, our results define a mechanism for SiRAV1 function in salt response where salt-triggered phosphorylation of SiRAV1 at Ser31 enhances its binding ability to SiCAT promoter, and the increased SiCAT expression contributes to salt tolerance in foxtail millet.

Key words: foxtail millet , salt stress , phosphoproteomic , SiRAV1 , SiCAT

LI Qiao-lu, LI Zhi-yong, WANG Meng-meng, YAN Jing-wei, FANG Lin. SiRAV1蛋白的Ser31位点磷酸化可以通过调节SiCAT表达来提高谷子耐盐性[J]. Journal of Integrative Agriculture, 2023, 22(12): 3638-3651.

LI Qiao-lu, LI Zhi-yong, WANG Meng-meng, YAN Jing-wei, FANG Lin. Phosphorylation of SiRAV1 at Ser31 regulates the SiCAT expression to enhance salt tolerance in Setaria italica[J]. Journal of Integrative Agriculture, 2023, 22(12): 3638-3651.

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